Skip to main content
SpringerLink
Log in

Detection of grass carp reovirus (GCRV) with monoclonal antibodies

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Grass carp reovirus (GCRV) is a pathogen that causes hemorrhagic disease of grass carp. It is the most serious infectious disease of carp and causes serious losses of fingerlings of grass carp and black carp. In this study, a recombinant VP4, one of the viral core proteins, was constructed with a histidine tag and expressed at a high level in E. coli, and the expressed protein was mainly found in the form of inclusion bodies. The expressed VP4 protein was recognized by an anti-His-tag monoclonal antibody and goat anti-GCRV serum. Four monoclonal antibodies (16B7, 39E12, 13C3 and 14D1) against the recombinant VP4 protein were produced. These MAbs did not react with any of the tested viruses or fish cells lines in the ELISA tests except GCRV. In western blotting analysis, a protein band was observed when the recombinant VP4 protein of GCRV was used as an antigen, but a 68-kDa band was observed when natural capsid proteins of GCRV were used as antigens. Furthermore, a sandwich ELISA was developed for detection of GCRV. The detection limit of the test was 105 TCID50 of GCRV per mL.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Jiang Y (2009) Hemorrhagic disease of grass carp: status of outbreaks, diagnosis, surveillance, and research. Israeli J Aquac Bamidgeh 61(3):188–197. http://evols.library.manoa.hawaii.edu/handle/10524/19287

    Google Scholar 

  2. Wolf K (1988) Fish viruses and fish viral diseases, pp 65–68

  3. Wang Q, Zeng W, Liu C, Zhang C, Wang Y, Shi C, Wu S (2012) Complete genome sequence of a reovirus isolated from grass carp, indicating different genotypes of GCRV in China. Virol J 86(22):12466

    Article  CAS  Google Scholar 

  4. Wang T, Li J, Lu L (2013) Quantitative in vivo and in vitro characterization of co-infection by two genetically-distant Grass carp reoviruses. J Gen Virol 94(Pt6):1301–1309

    Article  PubMed  CAS  Google Scholar 

  5. Ye X, Tian YY, Deng GC, Chi YY, Jiang XY (2012) Complete genomic sequence of a reovirus isolated from grass carp in China. Virus Res 163(1):275–283

    Article  PubMed  CAS  Google Scholar 

  6. Zhang C, Wang Q, Shi C, Zeng W, Liu Y, Wu S (2010) Molecular analysis of grass carp reovirus HZ08 genome segments 1–3 and 5–6. Virus Genes 41(1):102–104

    Article  PubMed  CAS  Google Scholar 

  7. Jiang Y, Ahne W (1989) Some properties of the etiological agent of the hemorrhagic disease of grass carp and black carp. In: Ahne W, Kurstak E (eds) Viruses of lower vertebrates. Springer, Berlin, pp 227–240

    Chapter  Google Scholar 

  8. Cheng L, Fang Q, Shah S, Atanasov IC, Zhou ZH (2008) Subnanometer-resolution structures of the grass carp reovirus core and virion. J Mol Biol 382:213–222

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  9. Cheng L, Zhu J, Hui WH, Zhang X, Honig B, Fang Q, Zhou ZH (2010) Backbone model of an aquareovirus virion by cry-electron microscopy and bioinformatics. J Mol Biol 397:852–863

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  10. Fang Q, Shah S, Liang Y, Zhou ZH (2005) 3D reconstruction and capsid protein character-ization of grass carp reovirus. Sci China C Life Sci 48(6):593–600

    Article  PubMed  CAS  Google Scholar 

  11. Fang Q, Seng EK, Ding QQ et al (2008) Characterization of infectious particles of grass carp reovirus by treatment with protease. Arch Virol 53:675–682

    Article  CAS  Google Scholar 

  12. Zhang X, Jin L, Fang Q, Hui WH, Zhou ZH (2010) 3 Å cryo-EM structure of a nonenveloped virus reveals a priming mechanism for cell entry. Cell 141(3):472–482

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  13. Andrew M, King Q, Michael J, Admas, Eric B (2012) Virus Taxonomy classification and nomenclature of viruses. In: Carstens, Lefkowitz EJ (eds) 9th Report of the International Committee on Taxonomy of Viruses, vol 1. Academic Press, San Diego, p 272

  14. Yan L, Guo H, Sun X, Shao L, Fang Q (2012) Characterization of grass carp reovirus minor core protein VP4. Virol J 9:89

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  15. Zhang QL, Yan Y, Shen JY, Hao GJ, Shi CY, Wang QT, Liu H, Huang J (2013) Development of a reverse transcription loop-mediated isothermal amplification assay for rapid detection of grass carp reovirus. J Virol Methods 187(2):384–389

    Article  PubMed  CAS  Google Scholar 

  16. Chi YY, Tian YY, Ye X, Deng GC, Li J, Wang HJ (2011) Molecular properties of grass carp reovirus in southern China and establishment of a duplex PCR detection method. Bing Du Xue Bao 27(4):358–365 (in Chinese)

    PubMed  CAS  Google Scholar 

  17. Zhang L, Luo Q, Fang Q, Wang Y (2010) An improved RT-PCR assay for rapid and sensitive detection of grass carp reovirus. J Virol Methods 169(1):28–33

    Article  PubMed  CAS  Google Scholar 

  18. Fijan ND, Sulimanovic M, Bearzotti D et al (1983) some properties of the epithelioma papulosum cyprini (EPC) cell line from carp (Cyprinus carpio). Ann Virol (Inst. Pasteur) 134E:207–220. http://www.sciencedirect.com/science/article/pii/S0769261783800604

  19. Gravel M, Malsbergen RG (1965) A permanent cell line from the fathead monnow (Pimephales promelas). Ann N Y Acad Sci 126:555–556

    Article  Google Scholar 

  20. Zuo WG, Qian HX, Xu YF et al (1984) A cell line derived from the kidney of grass carp (Ctenopharyngodon idellus). Fish China 2:38–39. http://en.cnki.com.cn/Article_en/CJFDTOTAL-SCKX198601001.htm

  21. Lannan CN, Winton JR, Fryer JL (1984) Fish cell lines: establishment and characterization of nine cell lines from salmonids. In Vitro 20:671–676

    Article  PubMed  CAS  Google Scholar 

  22. Wolf K, Quimby MC (1966) Lymphocystis virus: isolation and propagation in centrarchid fish cell lines. Science 151:1004–1005

    Article  PubMed  CAS  Google Scholar 

  23. Ahne W (1985) Studies on the use of fish tissue cultures for toxicity in order to reduce and replace the fish-tests. Zentralbl Bakteriol Mikrobiol Hyg B 180(5–6):480–504

    PubMed  CAS  Google Scholar 

  24. Wolf K, Quimby MC (1962) Established eurythermic line of fish cells in vitro. Science 135:1065–1066

    Article  PubMed  CAS  Google Scholar 

  25. Section of Virus Study, 3rd Laboratory, Institute of Hydrobiology, and Academia Sinica (1980) Studies on the causative agent of hemorrhage of the grass carp (Ctenopharyngodon idellus): electron microscopic observation. Acta Hydrobiol Sin 7:75–79

    Google Scholar 

  26. Ke LH, Fang Q, Cai Y (1990) Characteristics of a novel isolate of grass carp Hemorrhage Virus. Acta Hydrobiol Sinica 14:153–159 (in Chinese). http://en.cnki.com.cn/Article_en/CJFDTOTAL-SSWX199002006.htm

  27. Hoffmann B, Schütze H, Mettenleiter TC (2002) Determination of the complete genomic sequence and analysis of the gene products of the virus of Spring Viremia of Carp, a fish rhabdovirus. Virus Res 84(1–2):89–100

    Article  PubMed  CAS  Google Scholar 

  28. Zhang L-L, Shen J-Y, Lei C-F et al (2008) High level expression of Grass carp reovirus VP7 protein in prokaryotic cell. Virologica sinica 3(1):51–56. http://link.springer.com/content/pdf/10.1007%2Fs12250-008-2921-3

    Google Scholar 

  29. Shao L, Sun X, Fang Q (2011) Antibodies against outer-capsid proteins of grass carp reovirus expressed in E. coli are capable of neutralizing viral infectivity. Virol J 8:347

    Article  PubMed Central  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

We thank Prof. Yu-lin Jiang for critical reading of the revised manuscript. This research was supported by the National Science and Technology Pillar Program (2013BAD12B02) in China.

Author information

Authors and Affiliations

  1. Research Centre of Aquatic Animal Diseases, The Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China

    Jing Hongli, Zhang Min, Wang Na, Jiang Yulin & Lin Xiangmei

  2. Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing, 101113, China

    Zhang Lifeng

  3. Department of Fishery Sciences, Tianjin Agricultural University, Tianjin, 300384, China

    Fang Zhenzhen

  4. Beijing Fisheries Technology Extension Center, Beijing, 100021, China

    Xu Lipu

Authors
  1. Jing Hongli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhang Lifeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fang Zhenzhen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xu Lipu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhang Min
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wang Na
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiang Yulin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lin Xiangmei
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Jing Hongli or Lin Xiangmei.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hongli, J., Lifeng, Z., Zhenzhen, F. et al. Detection of grass carp reovirus (GCRV) with monoclonal antibodies. Arch Virol 159, 649–655 (2014). https://doi.org/10.1007/s00705-013-1864-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-013-1864-7

Keywords

Search

Navigation